The Optimization of Acquire Technology in the Medicinal Ingredients of Ganoderma Lucidiuma and Flammulina Velutipes
|School||Northeast Normal University|
|Keywords||Ganoderma Flammulina Immunomodulatory proteins Submerged fermentation Recombinant expression Physiological function|
Ganoderma lucidum is a well-known medicinal fungi, adjuvant therapy for tumor disease, chronic bronchitis, coronary heart disease, hepatitis, hyperlipidemia, neurasthenia, dermatomyositis diseases such as varying degrees of efficacy. Velutipes fungi both as a food and medicine, prevention and treatment of hepatitis, gastrointestinal ulcers, and has anti-cancer, reduce serum cholesterol. The traditional methods of training both fungi time-consuming, and the quality is not easy to control. With the development of modern science and production, and the submerged fermentation technology for large-scale production of these two fungi, with the production of large, small footprint, the cycle is short, easy to control the production process advantages. Immunomodulatory functions small proteins extracted from these two fungi - fungal immune regulatory proteins, having the biological activity of the agglutinated blood cells, also showed to stimulate lymphocyte proliferation, induction of immune cells to secrete a variety of cytokines and other immune regulator activity. This thesis is a preliminary study Ganoderma Flammulina submerged fermentation process. Two mycelium in the culture medium for different nitrogen source ratio peanuts and soybeans, the detection of two mycelia in shake flask and static culture growth conditions in the board of two different training methods and fresh and dry weight compare biological characteristics of the two kinds of culture mycelium in six different medium were determined. The test results show that the two stationary culture mycelium dry, fresh weight was significantly higher than that of the shake flask cultures; stationary culture, the total protein content of the mycelium peanut medium is slightly higher than that of soybean medium, the total sugar content of the basic equivalent to soybean medium, but liter peanut Medium cost than soybeans medium. Optimization of submerged fermentation the mycelia medium containing soybean flour 20g / L medium. To Pichia pastoris as host Flammulina recombinant expression of immunomodulatory proteins by phenol - sulfuric acid method analysis showed that the sugar content of pure recombinant protein reorganization fungal immunomodulatory protein glycosylation may occur in the yeast expression vector. After optimization of the induction of recombinant protein expression and purification technology, access to high expression and high purity the reorganization flammulina immunomodulatory protein. Of the purified recombinant flammulina immune regulatory proteins for the detection of biological activity, the results show that the recombinant protein aggregation activity of blood cells and promote the activity of the mouse lymphocyte proliferation. The paper submerged fermentation medium Ganoderma lucidum and flammulina liquid optimized, and also restructuring Flammulina immune optimized recombinant expression of regulatory proteins. Our results provide experimental basis, fungal immunomodulatory protein for further experimental study provides a theoretical basis for the production and development of the submerged fermentation of Ganoderma lucidum and Flammulina.